Center for Biomimetic Sensor Science, Nanyang Technological University, Singapore 637553, Singapore.
School of Materials Science and Engineering, Nanyang Technological University, Singapore 639798, Singapore.
Molecules. 2023 Feb 16;28(4):1909. doi: 10.3390/molecules28041909.
Exosomes released into the extracellular matrix have been reported to contain metabolic biomarkers of various diseases. These intraluminal vesicles are typically found in blood, urine, saliva, breast milk, cerebrospinal fluid, semen, amniotic fluid, and ascites. Analysis of exosomal content with specific profiles of DNA, microRNA, proteins, and lipids can mirror their cellular origin and physiological state. Therefore, exosomal cargos may reflect the physiological processes at cellular level and can potentially be used as biomarkers. Herein, we report an optical detection method for assaying exosomal biomarkers that supersedes the state-of-the-art time consuming and laborious assays such as ELISA and NTA. The proposed assay monitors the changes in optical properties of poly(3-(4-methyl-3'-thienyloxy) propyltriethylammonium bromide) upon interacting with aptamers/peptide nucleic acids in the presence or absence of target biomarkers. As a proof of concept, this study demonstrates facile assaying of microRNA, DNA, and advanced glycation end products in exosomes isolated from human plasma with detection levels of ~1.2, 0.04, and 0.35 fM/exosome, respectively. Thus, the obtained results illustrate that the proposed methodology is applicable for rapid and facile detection of generic exosomal biomarkers for facilitating diseases diagnosis.
已报道细胞外基质中释放的外泌体含有各种疾病的代谢生物标志物。这些腔囊泡通常存在于血液、尿液、唾液、母乳、脑脊液、精液、羊水和腹水。对具有特定 DNA、microRNA、蛋白质和脂质特征的外泌体内容物进行分析，可以反映其细胞起源和生理状态。因此，外泌体的货物可以反映细胞水平的生理过程，并可能被用作生物标志物。在此，我们报告了一种光学检测方法，用于检测外泌体生物标志物，该方法优于现有的耗时费力的检测方法，如 ELISA 和 NTA。该检测方法监测聚（3-（4-甲基-3'-噻吩氧基）丙基三乙基溴化铵）在存在或不存在靶标生物标志物的情况下与适体/肽核酸相互作用时的光学性质变化。作为概念验证，本研究证明了在人血浆中分离的外泌体中微 RNA、DNA 和晚期糖基化终产物的简便检测，检测水平分别约为 1.2、0.04 和 0.35 fM/外泌体。因此，所获得的结果表明，所提出的方法适用于快速简便地检测通用外泌体生物标志物，以促进疾病诊断。
